scholarly journals Effect of Sperm Cryopreservation on miRNA Expression and Early Embryonic Development

2021 ◽  
Vol 9 ◽  
Author(s):  
Xiaoyu Xu ◽  
Wanqiong Li ◽  
Lina Zhang ◽  
Yazhong Ji ◽  
Jiaying Qin ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Target Genes ◽  
Formation Rate ◽  
Fertilization Rate ◽  
Sperm Cryopreservation ◽  
Development Potential ◽  
Mouse Sperm ◽  
Negative Effect ◽  
Cryopreserved Sperm

Although sperm preservation is a common means of personal fertility preservation, its effects on embryonic development potential need further investigation. The purpose of this study was to identify key microRNA (miRNA) in cryopreserved sperm and determine the changes of these miRNAs and their target genes during embryonic development using cryopreserved sperm. Moreover, the embryonic development potential of cryopreserved sperm was estimated in assisted reproductive technology (ART), where key miRNAs and target genes were validated in sperm and subsequent embryos. Clinical data of embryonic development from cryopreserved sperm indicated a significant decrease in fertilization rate in both in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) cases, as well as a reduction in blastocyst formation rate in ICSI cases. Meanwhile there was a significant increase in blocked embryo ratio of Day1, Day2, and Day3.5 embryos when frozen-thawed mouse sperm was used, compared with fresh mouse sperm, suggesting a potential negative effect of sperm cryopreservation on embryonic development. From frozen-thawed and fresh sperm in humans and mice, respectively, 21 and 95 differentially expressed miRNAs (DEmiRs) were detected. miR-148b-3p were downregulated in both human and mouse frozen-thawed sperm and were also decreased in embryos after fertilization using cryopreserved sperm. Target genes of miR-148b-3p, Pten, was identified in mouse embryos using quantitative real-time PCR (qRT-PCR) and Western blot (WB). In addition, common characters of cryopreservation of mouse oocytes compared with sperm were also detected; downregulation of miR-148b-3p was also confirmed in cryopreserved oocytes. In summary, our study suggested that cryopreservation of sperm could change the expression of miRNAs, especially the miR-148b-3p across humans and mice, and may further affect fertilization and embryo development by increasing the expression of Pten. Moreover, downregulation of miR-148b-3p induced by cryopreservation was conserved in mouse gametes.

Effects of Different Types of Incubators on Embryo Development and Clinical Outcomes

2020 ◽  
Author(s):  
Ji Liu ◽  
Yan-Hua Zhou ◽  
Xiao-Xiao Wang ◽  
Ling-Xi Tong ◽  
Yan-Hong Li ◽  
...  
Keyword(s):  
Clinical Outcomes ◽  
Embryo Development ◽  
Culture Media ◽  
Controlled Trial ◽  
Formation Rate ◽  
Fertilization Rate ◽  
Vitro Fertilization ◽  
Water Jacket ◽  
Different Types

Abstract Background: Different types of incubators have been designed for gamete and embryo culture in the past few years. The main differences of these incubators are humidity, temperature and gas control system, which play important roles in regulating the steady state of culture media. The objective of this study was to compare the effects of different types of incubators (air jacket incubators and water jacket incubators) on embryo development and clinical outcomes in human in vitro fertilization (IVF).Methods: First, the physical performances of different incubators were tested by mimicking routine IVF procedures. After that, in a randomized controlled trial, 1013 cumulus oocyte complexes from 43 patients were equally divided into two groups, fertilized and cultured in two types of incubators to analyze the effects of different types of incubators on embryo development and clinical outcomes. Results: We found that temperature recovery time in the air jacket incubator was significantly shorter than that in water jacket incubator. Although the O2 recovering time was also significantly shorter in the air jacket incubator as compared with the water jacket incubator, no significant differences were observed in the CO2 recovering time between two groups, which was also verified by pH recovering time of culture media. Besides, the temperature of culture medium in the dish covered with oil recovered more quickly in the air jacket incubators than that in water jacket incubators. However, there were no significant differences observed in the fertilization rate, Day 3 high-quality embryo formation rate, blastocyst formation rate, good blastocyst rate and clinical outcomes between two groups.Conclusions: These results indicate that the microenvironment, especially the temperature, in air jacket incubator recover faster than that in conventional water jacket incubator, however, there were no significant differences in embryo development and clinical outcomes between two types of incubators.

208 EFFECT OF GDF8 AND SB-431542 ON PORCINE OOCYTE DURING IN VITRO MATURATION AND SUBSEQUENT EMBRYONIC DEVELOPMENT

2016 ◽  
Vol 28 (2) ◽  
pp. 235
Author(s):  
J. D. Yoon ◽  
E. Lee ◽  
S.-H. Hyun
Keyword(s):  
Treatment Group ◽  
Embryonic Development ◽  
Follicular Fluid ◽  
In Vitro Maturation ◽  
Transforming Growth Factor ◽  
Formation Rate ◽  
Transforming Growth Factor Β ◽  
Nuclear Maturation ◽  
Porcine Oocyte

Growth differentiation factor-8 (GDF8) is a member of the transforming growth factor-β that has been identified as a strong physiological regulator. SB-431542 (SB) is a specific inhibitor of transforming growth factor-β superfamily type I activin receptor-like kinase (ALK) receptors such as ALK4, ALK5, and ALK7. The purpose of this study is investigation of the effects of GDF8 and SB on porcine oocytes during in vitro maturation and subsequent embryonic development. We first performed ELISA to detect GDF8 concentrations in follicular fluid for each size of follicle; sizes were as follows: small (<3 mm), medium (>3 mm and <6 mm), and large (>6 mm) follicle. After detection of the GDF8 concentration in follicular fluid, we investigated the effect of GDF8 and SB treatment during in vitro maturation (IVM) on nuclear maturation, intracellular glutathione (GSH), and reactive oxygen species (ROS) levels, and embryonic development after IVF and parthenogenetic activation (PA). Data were analysed by ANOVA followed by Duncan using SPSS (Statistical Package for Social Science, IBM, New York, NY, USA) mean ± SEM. The ELISA result showed different concentrations of GDF8 for each grade of follicular fluid: small, 0.479 ng mL–1; medium, 0.668 ng mL–1; and large, 1.318 ng mL–1. During the IVM process, 1.318 ng mL–1 of GDF8 and 5 ng mL–1 of SB were added to the maturation medium as control, SB, SB+GDF8, and GDF8 treatment groups. After 44 h of IVM, GDF8 group (90.4%) showed a significantly higher nuclear maturation rate than control and SB+GDF8 groups (85.4 and 81.7%). The SB group (78.9%) showed significantly reduced nuclear maturation rate compared with control (P < 0.05). The GDF8 treatment group showed a significant decreased intracellular ROS and increased GSH levels compared with other groups (P < 0.05). The SB+GBF8 treatment group showed a significantly better cytoplasmic maturation than the SB treatment group. In the PA embryonic development analysis, the GDF8 treatment group showed a significantly higher blastocyst formation rate compared with other groups (47.9, 37.2, 46.4, and 58.7% respectively; P < 0.05). In the IVF embryonic development analysis, the GDF8 treatment groups showed significantly higher blastocyst formation rate compared with the SB group (28.2 and 42.2%, respectively; P < 0.05). In conclusion, treatment with GDF8 during porcine oocyte IVM improved the embryonic developmental competence via increased cytoplasmic maturation and led to better oocyte maturation from the ALK receptor inhibition by SB.

137 Kinetics of In Vitro Embryonic Development According to the Follicular Population in Bos Indicus

2018 ◽  
Vol 30 (1) ◽  
pp. 208
Author(s):  
J. G. Soares ◽  
F. M. Morato ◽  
G. F. Rossi ◽  
B. M. Bayeux ◽  
A. S. Oliveira ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Formation Rate ◽  
Bos Indicus ◽  
Prostaglandin F2α ◽  
Lower Percentage ◽  
Embryo Production ◽  
High Category ◽  
Kinetics Of ◽  
In Vitro Embryonic Development

The present study aimed to evaluate the effect of the follicular population from cull Nelore (Bos indicus) on the kinetics of in vitro embryonic development. At random stages of the oestrous cycle (Day –5), a total of 28 cull Nelore cows were synchronized with an intravaginal progesterone device associated with oestradiol benzoate (2.0 mg IM). At the same moment, a dose of prostaglandin F2α (2.0 mg im) was also administered to promote luteolysis and absence of corpus luteum (CL) at the time of ovum pick-up (OPU). Five days later (Day 0), all cows underwent an OPU session and the recovered oocytes were submitted to in vitro embryo production (IVEP). The same procedures were repeated 2 times at 30-day intervals (Day 25 and Day 55). Semen from a single batch of a previously tested bull was used for all IVEP. Blastocyst production and hatching were verified on Days 7, 8, and 9 of the IVEP. Data were analysed by the GLIMMIX procedure of SAS 9.3® (SAS Institute Inc., Cary, NC, USA). Data of the 3 OPU sessions were grouped and the cows were classified into 3 categories according to the follicular population: Low (19.7 ± 0.9 follicles, n = 27), Medium (33.5 ± 0.8 follicles, n = 29), and High (58.7 ± 3.2 follicles, n = 28). The Low category had a lower rate of viable oocytes [(number of viable oocytes/total number of oocytes) × 100; 62.0 v. 69.5%; P = 0.02] and cleavage rate [(number of cleaved/total number of oocytes) × 100; 55.9 v. 66.8%; P = 0.001] than the High category. The blastocyst formation rate [number of blastocysts/total number of oocytes) × 100] on Day 7 and Day 8 was lower for Low and Medium compared with the High category (Day 7: 26.1b v. 29.0b v. 35.1a %; P = 0.001; and Day 8: 29.2b v. 30.2b v. 34.7a %; P = 0.05). No differences were found in blastocysts rate on Day 9 among Low, Medium, and High categories (14.1 v. 15.9 v. 16.2%; P = 0.61). However, Low category had a lower percentage of hatched blastocysts [(number of hatched blastocysts/number of blastocysts) × 100] on Day 7 compared with High category (2.9 v. 12.0%; P = 0.01). These results reported that cull Nelore with High follicular population showed higher rates of embryo production and hatched blastocysts compared with cows with Low follicular population. We concluded that the kinetics of in vitro embryonic development was compromised in cull Nelore (Bos indicus) with low follicular population submitted to OPU-IVEP. This research was supported by Fapesp 2012/50533–2 (GIFT).

Study of Morphokinetics in Day 3 Embryo with Implantation Potential and Effect of Sperm Cryopreservation on Embryogenesis

2017 ◽  
Vol 8 (2) ◽  
pp. 61-67
Author(s):  
Harsha K Bhadarka ◽  
Nayana H Patel ◽  
Kruti B Patel ◽  
Nilofar R Sodagar ◽  
Yuvraj D Jadeja ◽  
...  
Keyword(s):  
Time Lapse ◽  
Mean Value ◽  
Primary Objective ◽  
Sperm Cryopreservation ◽  
Cell Stage ◽  
Frozen Semen ◽  
Significant Difference ◽  
Cryopreserved Sperm ◽  
Day 3 Embryo

ABSTRACT Aim In recent past, many studies had come up with the combination of time-lapse (TL) imaging of embryo morphokinetics as a noninvasive means for improving embryo selection and in vitro fertilization (IVF) success. The primary objective of the study was to find out if there is significant variation in morphokinetics of embryos with different implantation potential and also to study the effect of sperm freezing on time points of embryogenesis events in embryos with implantation potential. Materials and methods Kinetic data and cycle outcomes were analyzed retrospectively in 142 patients who had undergone IVF/intracytoplasmic sperm injection (ICSI) cycles using semen with normal parameters and embryo transfer (ET) on day 3. For the surety of specificity of morphokinetics, only cases with single ET cycles were included in the study. Timing of specific events, from the point of ICSI, was determined using TL imaging. Kinetic markers like time to syngamy (t-pnf), t2, time to two cells (c), 3c (t3), 4c (t4), 5c (t5), 8c (t8), tMor, CC2, CC3, t5–t2, t5–t4, s1, s2, and s3 were calculated. The cleavage synchronicity from the 2–8 cell stage (CS2–8), from 4 to 8 cell stage (CS4–8), and from 2 to 4 cell stage (CS2–4) were calculated as defined elsewhere. Deoxyribonucleic acid replication time ratio (DR) was also included in the comparison. Analysis of variance test was used for comparison of the mean timing of cell division and cell cycle intervals. Results Morphokinetics t-pnf, t2, t8, CC2, S2, S3, CS2–8, CS4–8, and CS2–4 differed significantly between embryos with and without implantation potential, when embryos were developed using fresh semen, while t3, t4, t5, CC2, S2, t5–t2, CS2–4, and DR differed significantly between the embryos with and without implantation potential when frozen semen was used. No significant difference was found in mean value of any of the above-stated parameters when comparison was done between implanted embryos fertilized by either fresh or cryopreserved sperm. Conclusion Many morphokinetics parameters of embryogene­sis vary significantly between embryos with different ability to implant; therefore, the criteria developed in our IVF lab can be useful for selection of suitable embryo even at day 3 of development with more chances of implantation. Clinical significance Study indicates necessity of development of individualized selection model based on morphokinetics for every IVF lab and also confirms freezing as an important tool for fertility preservation of males as it does not affect events of embryogenesis. How to cite this article Bhadarka HK, Patel NH, Patel KB, Sodagar NR, Jadeja YD, Patel NH, Patel MN, Patel AV, Patel DH, Patel JS. Study of Morphokinetics in Day 3 Embryo with Implantation Potential and Effect of Sperm Cryopreservation on Embryogenesis. Int J Infertil Fetal Med 2017;8(2):61-67.

scholarly journals A Method for Chronological Intravital Imaging of Bovine Oocytes during In Vitro Maturation

2008 ◽  
Vol 14 (6) ◽  
pp. 549-560 ◽  
Author(s):  
Morten R. Petersen ◽  
Michael Hansen ◽  
Birthe Avery ◽  
Ingrid B. Bøgh
Keyword(s):  
Embryonic Development ◽  
Oocyte Maturation ◽  
Laser Scanning ◽  
Subsequent Development ◽  
Cumulus Cells ◽  
Fertilization Rate ◽  
Laser Scanning Microscopy ◽  
Intravital Imaging ◽  
Bovine Oocytes

AbstractOocyte maturation is known to affect the chances for successful fertilization, embryonic development, establishment of pregnancy and delivery of a live, healthy, and viable offspring. Two-photon laser scanning microscopy (TPLSM) has previously been used to evaluate early embryonic development without a detectable impairment of subsequent development, but has never been applied to assess mammalian oocytes throughout in vitro maturation (IVM). Visualization of structures within live oocytes during IVM, followed by fertilization and embryo culture, may improve the understanding of oocyte maturation. To visualize structures within bovine oocytes using TPLSM, it is necessary to remove the cumulus cells that normally surround the oocyte during maturation. Repeated visualization of structures within the same oocyte is possible, if movement of the oocyte can be avoided. In this article, we describe the development of a method for repeated intravital imaging of denuded bovine oocytes using an upright TPLSM equipped with a specially constructed incubator. Oocytes were stained with Hoechst 33258, and the nuclear structures were evaluated. Oocyte fertilization rate was not affected by TPLSM exposure, but the developmental capacity of the denuded oocytes was significantly reduced. This is, to our knowledge, the first article describing repeated intravital imaging during mammalian oocyte maturation using TPLSM.

scholarly journals Effects of cryopreservation on sperm quality, nuclear DNA integrity, in vitro fertilization, and in vitro embryo development in the mouse

Reproduction ◽  
2007 ◽  
Vol 133 (3) ◽  
pp. 585-595 ◽  
Author(s):  
Cengiz Yildiz ◽  
Palma Ottaviani ◽  
Napoleon Law ◽  
Renise Ayearst ◽  
Ling Liu ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Embryo Development ◽  
Membrane Integrity ◽  
Fertilization Rate ◽  
Development Rate ◽  
Dna Integrity ◽  
Plasma Membrane Integrity ◽  
Mouse Sperm ◽  
Progressive Motility

Efficient freezing, archiving, and thawing of sperm are essential techniques to support large scale research programs using mouse models of human disease. The purpose of this study was to investigate the effects of variable combinations and concentrations of cryoprotectants on sperm-assessment parameters of frozen–thawed mouse sperm in order to optimize cryopreservation protocols. Sperm was frozen using combinations of 3% skim milk + 0.2 or 0.3 M nonpermeating raffinose with either permeating glucose, fructose, propylene glycol, ethylene glycol, glycerol, or sodium pyruvate in CD-1, C3FeB6F1/J, B6129SF1, C57BL/6NCrIBR, 129S/SvPaslco, and DBA/2NCrIBR mice. Sperm-assessment parameters included progressive motility, plasma membrane integrity (SYBR-14 + PI),in vitrofertilization rate, andin vitroembryo development rate to blastocyst. DNA content analysis of sperm was measured by the sperm chromatin structure assay (SCSA). 0.3 M raffinose with 0.1 M fructose significantly improved post-thaw sperm-assessment parameters for CD-1, C3B6F1, B6129SF1 mice (P< 0.05–0.01), whereas 0.2 M raffinose with 0.1 M glycerol or 0.1 M fructose enhanced sperm assessment values for C57BL/6 and 129S mice (P< 0.01), compared to 0.3 M raffinose alone. DNA fragmentation during cryopreservation was significantly increased in all strains evaluated when compared with fresh control sperm in a strain-dependent manner (P< 0.01). Supplementation with permeating glycerol or fructose to the cryoprotectant (CPA) solution showed a significant protective effect to DNA integrity when cryopreserving sperm from C57BL/6 and 129S mice. Damage to sperm DNA significantly decreased the rate ofin vitroembryo development to blastocyst in C57BL/6 mice. The type of monosaccharide sugar or polyols, CPA molarity, and combination of permeating and nonpermeating cryoprotectant are significant factors for improving progressive motility, plasma membrane integrity, DNA integrity,in vitrofertilization rate, andin vitroembryo development rate to blastocyst in cryopreserved mouse sperm.

scholarly journals Sperm DNA damage has a negative effect on early embryonic development following in vitro fertilization

2018 ◽  
Vol 20 (1) ◽  
pp. 75 ◽  
Author(s):  
Wei-Wei Zheng ◽  
Ge Song ◽  
Qi-Ling Wang ◽  
Shan-Wen Liu ◽  
Xiao-Li Zhu ◽  
...  
Keyword(s):  
Dna Damage ◽  
In Vitro Fertilization ◽  
Embryonic Development ◽  
Early Embryonic Development ◽  
Vitro Fertilization ◽  
Sperm Dna Damage ◽  
Sperm Dna ◽  
Negative Effect

scholarly journals Penambahan Glutathione pada Medium Fertilisasi Efektif Mendukung Pembentukan Pronukleus dan Perkembangan Awal Embrio Sapi (SUPPLEMENTATION OF GLUTATHIONE IN FERTILIZATION MEDIUM EFFECTIVELY SUPPORT NORMAL PRONUCLEUS FORMATION AND EARLY BOVINE EMBRYONIC DE

2017 ◽  
Vol 18 (3) ◽  
pp. 327
Author(s):  
Aras Prasetiyo Nugroho ◽  
Iman Supriatna ◽  
Mohamad Agus Setiadi
Keyword(s):  
Tissue Culture ◽  
Embryonic Development ◽  
Culture Medium ◽  
Fertilization Rate ◽  
Tissue Culture Medium ◽  
Randomized Design ◽  
Oviduct Fluid ◽  
And Control ◽  
Bovine Oocytes

The objective of this study was to determine fertilization rate effectiveness and early embryonic development competency with glutathione (GSH) supplementation in fertilization medium and culture This study consisted of two experiments comprising each of the four treatment and six repetitions with completely randomized design (CRD) using 651 oocytes. In the first experiment, a total of 317 bovine oocytes were matured in tissue culture medium (TCM) 199 at incubator 5% CO2 with temperature 39 ºC for24 h, then fertilized with sperm separated by swim up technique. Oocyte and sperms were incubated in fertilization medium supplemented with 0.25 mM, 0.50 mM, 1.00 mM GSH. In the second experiment, bovine oocytes were matured in maturation medium and fertilized with same procedure as mentioned before, then cultured in modified synthetic oviduct fluid (mSOF) with the following treatment: supplementation GSH only in fertilization medium (T1), supplementation GSH only in culture medium (T2), and supplementation GSH in both fertilization and culture medium (T3), while control not supplementation GSH (T0). Result of the first experiment showed that supplementation 1.00 mM GSH in fertilization medium can increase higher normal pronucleus (2PN) formation (86,9%) compared to other treatments, 0.50 mM (80.3%), 0.25 mM (73.8%), and control (58.9%) (P<0.05). In the second experiment showed that early bovine embryonic development on 2nd day cultured which reached 5-8 cell on treatment T1 (56.0%) and T3 (53.6%) were higher (P<0.05) compared to treatment T2 (26.2%) and T0 (control) (31.3%). Result of the other were showed that early bovine embryonic development on 4th day cultured which reached 9-16 cell on treatment T1 (26.2%) and T3 (27.4%) were higher (P<0.05) compared to that T2 (11.9%) and T0 (control) (10.8%). In conclusion, 1.00 mM GSH supplementation in medium was more effective in supporting normal pronucleus formation and early fertilization bovine embryonic development compared to in culture medium. ABSTRAK Penelitian ini bertujuan untuk mengetahui tingkat fertilisasi dan kompetensi perkembangan awal embrio sapi dengan penambahan glutathione (GSH) pada medium fertilisasi in vitro (IVF) dan kultur in vitro (IVC). Penelitian ini terdiri atas dua penelitian yang terdiri dari masing-masing empat perlakukan dan enam kali ulangan dengan rancangan acak lengkap (RAL) menggunakan 651 oosit. Penelitian I, sebanyak 317 oosit sapi dalam tissue culture medium (TCM) 199 dimatangkan pada inkubator 5% CO2 dan suhu 39°C selama 24 jam, kemudian difertilisasi dengan spermatozoa yang telah diseleksi menggunakan teknik swim up. Oosit dan spermatozoa diinkubasi pada medium fertilisasi dengan penambahan 0,25 mM, 0,50 mM, dan 1,00 mM GSH. Penelitian II, sebanyak 334 oosit sapi dimatangkan pada medium pematangan dan difertilisasi, kemudian dikultur pada medium modified synthetic oviduct fluid (mSOF), dengan perlakuan: penambahan GSH hanya pada medium fertilisasi (T1), penambahan GSH hanya pada medium kultur (T2), dan kombinasi penambahan GSH pada medium fertilisasi dan kultur (T3). Hasil penelitian I, menunjukkan bahwa penambahan 1,00 mM GSH pada medium fertilisasi dapat meningkatkan pembentukan pronukleus normal (2PN) yang lebih tinggi (86,9%) dibandingkan dengan perlakuan yang lain yaitu 0,50 mM (80,3%), 0,25 mM (73,8%), dan kontrol (58,9%) (P<0,05). Penelitian II menujukkan bahwa perkembangan awal embrio sapi pada hari ke-2 kultur yang mencapai pembelahan 5-8 sel pada perlakukan T1 (56,0%) dan T3 (53,6%) lebih tinggi (P<0,05) dibandingkan dengan perlakuan T2 (26,2%) dan T0 (kontrol) (31,3%). Hasil penelitian lain menunjukkan bahwa perkembangan awal embrio sapi pada hari ke-4 kultur yang mencapai pembelahan 9-16 sel pada perlakuan T1 (26,2%) dan T3 (27,4%) lebih tinggi dibandingkan dengan perlakukan T2 (11,9%) dan T0 (kontrol) (10,8%) (P<0,05). Dapat disimpulkan bahwa penambahan 1,00 mM GSH pada medium fertilisasi lebih efektif dalam mendukung pembentukan pronukleus normal dan perkembangan awal embrio sapi dibandingkan pada medium kultur.

200 SYNCHRONIZATION OF OOCYTE MEIOTIC MATURATION IN SUPEROVULATED MICE IMPROVES IN VITRO FERTILIZATION RATE

2012 ◽  
Vol 24 (1) ◽  
pp. 212
Author(s):  
A. M. Taiyeb Ridha ◽  
D. C. Kraemer
Keyword(s):  
Embryonic Development ◽  
Fertilization Rate ◽  
Early Embryonic Development ◽  
Meiotic Maturation ◽  
Control Group ◽  
Vitro Fertilization ◽  
Development Rates ◽  
Oocyte Meiotic Maturation

In vitro synchronization of oocyte nuclear and cytoplasmic maturation has been found to improve the IVF rate of ovarian oocytes in several species, including humans, in comparison with nonsynchronized in vitro-matured oocytes. Here, we tested the hypothesis that synchronization of oocyte meiotic maturation by an in vivo system in superovulated mice would increase the oocyte fertilization rate when compared to that of conventional superovulated oocytes. Recently, we observed that cilostazol (CZL), a PDE3-I, was able to inhibit mouse oocyte meiotic maturation in both in vitro and in vivo systems. Administering CZL at 7.5 mg, 4 or 7 h pre-hCG allowed retrieval of ovulated oocytes of which >95% were at MI stage, scored by Nikon stereo microscope (SMZ 1500). A conventional superovulation program was adapted in all treated and their control groups, in which mice were injected with eCG and after 48 h with hCG (7.5 IU for each hormone). On the second morning, 13 to 14 h post-hCG, mice were killed and oocytes were collected from oviducts and in vitro fertilized (control). For the treated groups, CZL was administered in a single 7.5 mg oral dose (gavage) 4 or 7 h before the hCG injection. On the second morning, CZL-treated animals were killed at the same timing as control animals and oocytes were retrieved from the oviduct and in vitro matured for 6 h (for those gavaged with CZL, 4 h pre-hCG) or 3 h (for those gavaged with CZL, 7 h pre-hCG) to MII oocytes before IVF. These groups were designated as in vivo-in vitro synchronized/matured oocytes. In other groups treated with CZL, 4 or 7 h pre-hCG, the ovulated oocytes were allowed to mature in the oviduct (full in vivo synchronization and maturation) and oocytes were retrieved and fertilized with the same fertilization timings as the in vivo-in vitro synchronized/matured oocytes. Oocytes were cultured for 1 day after IVF and examined for cleavage. Statistical differences were analyzed by cross-tabulated chi-square test. The full in vivo synchronization and maturation (for both CZL dose timings of 4 and 7 h pre-hCG) gave significantly higher early embryonic development rates compared with those of the control [89% (n = 219) and 92.2% (n = 374) vs 81.8% (n = 198); P = 0.034 and P < 0.0001, respectively]. The in vivo-in vitro synchronized/matured oocytes (CZL dose timing at 7 h, but not 4 h pre-hCG) gave significantly higher early embryonic development rates compared with those of the control [88.5% (n = 339) vs 83.4% (n = 458), respectively; P = 0.043]. However, the increase of the IVF rate of the oocytes from mice treated with CZL, 4 h pre-hCG, in the in vivo-in vitro synchronized/matured group was not significantly different from the control group [88.5% (n = 399) vs 83.4% (n = 458), respectively; P = 0.43]. It is concluded from the present study that synchronization of oocyte meiotic maturation by the in vivo and in vivo-in-vitro protocols can increase the IVF rate of oocytes in superovulated mice.

Fertilization and early embryonic development of in vitro matured metaphase I oocytes in patients with unexpected low oocyte maturity rate

Zygote ◽  
2021 ◽  
pp. 1-5
Author(s):  
Nafiye Yılmaz ◽  
Şebnem Özyer ◽  
Derya Taş ◽  
Mehmet Caner Özer ◽  
Ayten Türkkanı ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Fertilization Rate ◽  
Early Embryonic Development ◽  
Oocyte Maturity ◽  
Significant Difference ◽  
Maturation Rate ◽  
Developmental Capacity ◽  
Metaphase I

Summary To determine the fertilization and embryonic potential of immature metaphase I (MI) oocytes in patients with low oocyte maturity rate in whom the percentage of mature oocytes obtained was less than 75% of the total retrieved ones. In vivo matured metaphase II (MII) oocytes (MII-ICSI, n = 244), and in vitro matured MI oocytes (MI-MII-ICSI, n = 202) underwent an intracytoplasmic sperm injection (ICSI) procedure. Maturation rate, fertilization rate and early embryonic development were compared in both groups. In total, 683 oocytes were collected from 117 ICSI cycles of 117 patients. Among them, 244 (35.7%) were mature MII and 259 (37.9%) were MI after the denudation process. Of those 259 MI oocytes, 202 (77.9%) progressed to MII oocytes after an incubation period of 18–24 h. The maturation rate was 77.9%. Fertilization rate was found to be significantly higher in the rescued in vitro matured MI oocyte group when compared with the in vivo matured MII oocyte group (41.6% vs 25.8%; P = 0.0006). However, no significant difference was observed in terms of cleavage rates on days 2 and 3 between the groups (P = 0.9126 and P = 0.5031, respectively). There may be unidentified in vivo factors on the oocyte maturation causing low developmental capacity in spite of high fertilization rates in the group of patients with low oocyte maturity rate. Furthermore, studies are needed to determine the appropriate culture characteristics as well as culture period and ICSI timing of these oocytes.

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